CN107748263A - The new application and kit of a kind of plasminogen - Google Patents
The new application and kit of a kind of plasminogen Download PDFInfo
- Publication number
- CN107748263A CN107748263A CN201710775306.8A CN201710775306A CN107748263A CN 107748263 A CN107748263 A CN 107748263A CN 201710775306 A CN201710775306 A CN 201710775306A CN 107748263 A CN107748263 A CN 107748263A
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- Prior art keywords
- plg
- kit
- assessment
- parts
- tbi
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Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
Abstract
The present invention relates to a kind of new application of plasminogen and kit., can accurate judgement TBI patient injury's orders of severity by detecting the content of PLG in cerebrospinal fluid；The kit prepared by above method, the content of PLG in cerebrospinal fluid can be effectively detected, and then accurately assessed or aided assessment TBI patient injury's orders of severity, have that specificity is good, susceptibility is high, high repeatability and other advantages, there is very big application value.
Description
Technical field
The invention belongs to medical field, the new application and kit of more particularly to a kind of plasminogen.
Background technology
Characters of traumatic brain injury (TBI) is a key factor for influenceing human health in the world, has high generation
Rate, there are 2,000,000 TBI in statistics display, the U.S. every year recently, wherein 43% TBI patient can leave deformity, China is every
There are 1,000,000 TBI in year, and Chinese head injury data are shown, the death rate of TBI patient can reach 21.8%.
According to degree of injury, TBI can be divided into slight, moderate and severe TBI.Its division methods Main Basis Ge Lasi
Brother's Coma Score (GCS), i.e., in TBI, patients of the GCS between 3 to 8 is the patient of severe TBI, GCS between 9 to 12
For moderate TBI, patients of the GCS between 13 to 15 is slight TBI.Wherein GCS scorings are mainly anti-by reaction of opening eyes, language
TBI injury in traumatic condition of patient should be assessed with motor reaction, due to reasons such as the experiences of the doctor in charge, assessment result often has
Certain subjectivity, and TBI is usually associated with multiple injury, causes doctor according to the eye opening unrelated with TBI, language and motion
Erroneous judgement of the obstacle to the TBI state of an illness, in addition, the language of country variant, different geographical also likely to be present certain communication disorder.
The content of the invention
In order to solve the above-mentioned technical problem, the invention provides a kind of new application of plasminogen and kit,
The new application and kit can be used for assessing the TBI injury in traumatic condition of patient orders of severity, and result is accurate, easy to operate, have larger
Application value.
Concrete technical scheme of the present invention is as follows：
The invention provides the material for detecting plasminogen (PLG) to prepare assessment or aided assessment TBI
Application in the product of patient injury's order of severity.
The invention provides the material for detecting PLG to prepare assessment or aided assessment TBI patient injury's orders of severity
Kit in application.
By the content for detecting PLG in cerebrospinal fluid, it can be estimated that or the badly wounded degree of aided assessment TBI patient, should
Appraisal procedure specificity and sensitivity are high, favorable reproducibility.
Further improve, the antibody that the material for detecting PLG is anti-PLG.
Preferably, PLG antibody specific is rabbit-anti PLG antibody.
It is further preferred that rabbit-anti PLG antibody is the rabbit-anti PLG antibody of abcam companies.
Further, PLG sequence is as shown in SEQ ID NO.1.
Present invention also offers applications of the PLG in the label for assessing the TBI injury in traumatic condition of patient orders of severity is prepared.
Present invention also offers a kind of kit for being used for assessment or aided assessment TBI patient injury's orders of severity, the examination
Agent box includes being used for the material for detecting PLG.
The present invention, can be by detecting PLG in cerebrospinal fluid by adding the material for detecting PLG in kit
Content, assessment or aided assessment TBI patient injury's orders of severity.
Further to improve, kit includes preserving liquid, trehalose, the 15-30 for preserving liquid by parts by weight for 1-5 parts
The 1640 culture medium composition of the glycerine of part, the serum of 20-30 parts and 10-20 parts.
The present invention is by using trehalose, glycerine, serum and 1640 culture medium as the constituent for preserving liquid, making enzyme mark
The holding time of anti-PLG antibody is longer, keeps the activity of the anti-PLG antibody of enzyme mark so as to PLG in more accurate measure cerebrospinal fluid
Content, draw definite diagnostic result.
Further improve, kit also includes dilution, the dilution by 50-70 parts phosphate buffer
(PBS), the tween 100 (tween100) of 10-25 parts and the bovine serum albumin(BSA) (BSA) of 1-5 parts composition.
The present invention makes the anti-PLG antibody of enzyme mark by the constituent using PBS, tween100 and BSA as dilution
Dilution effect is more preferable, so as to preferably determine its PLG content.
Content of the invention by detecting PLG in cerebrospinal fluid, can accurate judgement TBI patient injury's orders of severity；By with
Kit prepared by top method, it can effectively detect the content of PLG in cerebrospinal fluid, and then accurately assessment or aided assessment TBI
Patient injury's order of severity.
Brief description of the drawings
Fig. 1 is the result of the test of test example 1, and Fig. 1 is that TBI rat cerebral tissues damage zone and rats in sham-operated group are corresponding
The fluorogram of position albumen；1 represents rabbit-anti PLG antibody mediated immunity fluorescence analysis figures, and 2 represent to redye chromatic graph with DAPI；In 3
Merged represents a, b fusion figure；
Fig. 2-6 is the result of the test of test example 2, and Fig. 2 is the PLG testing results of rabbit-anti PLG antibody；Fig. 3 is the anti-PLG of mouse
The PLG testing results of antibody；Fig. 4 is the PLG testing results of goat-anti PLG antibody；Fig. 5 is the limited public affairs of the auspicious neat life science in Shanghai
Take charge of the PLG testing results of rabbit-anti PLG antibody；Fig. 6 is the PLG of the Bioisystech Co., Ltd of Pu Xin containing Shanghai rabbit-anti PLG antibody
Testing result.
Embodiment
Embodiment 1
A kind of kit, including liquid is preserved, wherein preserving liquid includes the composition of following weight：
1 part of trehalose, 15 parts of glycerine
20 parts of serum, 10 parts of 1640 culture medium.
Embodiment 2
A kind of kit, including liquid is preserved, wherein preserving liquid includes the composition of following weight：
3 parts of trehalose, 25 parts of glycerine
25 parts of serum, 15 parts of 1640 culture medium.
Embodiment 3
A kind of kit, including liquid is preserved, wherein, preserving liquid includes the composition of following weight：
5 parts of trehalose, 30 parts of glycerine
30 parts of serum, 20 parts of 1640 culture medium.
Embodiment 4
A kind of kit, including dilution, wherein, dilution includes the composition of following weight：
50 parts of PBS, 10 parts of tween100
1 part of BSA.
Embodiment 5
A kind of kit, including dilution, wherein, dilution includes the composition of following weight：
60 parts of PBS, 15 parts of tween100
3 parts of BSA.
Embodiment 6
A kind of kit, including dilution, wherein, dilution includes the composition of following weight：
70 parts of PBS, 25 parts of tween100
5 parts of BSA.
Reference examples 1
A kind of kit, including liquid is preserved, wherein, preserving liquid includes the composition of following weight：
1 part of trehalose, 15 parts of glycerine
10 parts of serum, 10 parts of 1640 culture medium
10 parts of citric acid.
Reference examples 2
A kind of kit, including liquid is preserved, wherein, preserving liquid includes the composition of following weight：
1 part of trehalose, 15 parts of glycerine
20 parts of citric acid, 10 parts of 1640 culture medium.
Reference examples 3
A kind of kit, including liquid is preserved, wherein, preserving liquid includes the composition of following weight：
1 part of trehalose, 15 parts of glycerine
20 parts of serum.
Reference examples 4
A kind of kit, including dilution, wherein, dilution includes the composition of following weight：
25 parts of PBS, 10 parts of tween100
1 part of BSA, 25 parts of sodium sulphate.
Reference examples 5
A kind of kit, including dilution, wherein, dilution includes the composition of following weight：
25 parts of PBS, 10 parts of tween100
26 parts of sodium sulphate.
Reference examples 6
A kind of kit, including dilution, wherein, dilution includes the composition of following weight：
50 parts of PBS, 11 parts of BSA.
The plasminogen of test example 1 can be as the discovery procedure of the label of the TBI injury in traumatic condition of patient orders of severity
Paired sample：6 TBI SD rats, and sham-operation SD rats.
TBI rat cerebral tissues damage zone and rats in sham-operated group corresponding position albumen are marked by iTRAQ,
Using Damage of Rats area brain after quantitative proteomics UPLC-LTQ Orbitrap-Velos ultrahigh pressure liquid phases and mass spectral analysis TBI
Histone expression change；MaxQuant (version 1.5.3.28) search Swiss-Prot (version release 2015_08,
7928query number) database.Classical calorific power：Protein abundance increases by 1.0 times or protein abundance reduces 1.0 times.
Confirm that PLG genes expression quantity in TBI groups significantly raises by immunofluorescence technique.Specific experimental method is such as
Under:Each sample is taken respectively, carries out Immunohistochemical detection.Primary antibody is PLG antibody (Abcam, article No.：Ab154560), two
Resist for goat-anti rabbit fluorescence secondary antibody (Abcam, article No.：Ab150077), Fig. 1 is as a result seen.
Level differences of the test example 2PLG in TBI Cerebrospinal Fluid in Patients
Detect sample：The cerebrospinal fluid (55 slight TBI, 61 moderates TBI, 62 severe TBI) of 178 TBI patients.
Wherein, each patient is the volunteer of informed consent.
Packet：
First group：The assessment of the rabbit-anti PLG Antibody preparations of abcam companies or aided assessment TBI patient injury's orders of severity
Product.
Second group：The assessment of the anti-PLG Antibody preparations of mouse of abcam companies or aided assessment TBI patient injury's orders of severity
Product.
3rd group：The assessment of the goat-anti PLG Antibody preparations of abcam companies or aided assessment TBI patient injury's orders of severity
Product.
4th group：The assessment of the rabbit-anti PLG Antibody preparations of Shanghai Rui Qi life sciences Co., Ltd or aided assessment TBI
The product of patient injury's order of severity.
5th group：The assessment of the rabbit-anti PLG Antibody preparations of Shanghai Pu Xin Bioisystech Co., Ltd or aided assessment TBI
The product of patient injury's order of severity.
Each sample is taken respectively, carries out enzyme-linked immunosorbent assay (enzyme-linked with five set respectively
Immunosorbent assay, ELISA) detect, determine PLG content, 178 TBI Cerebrospinal Fluid in Patients PLG testing results point
Do not see Fig. 2-6 (each point represents 1 sample).
From Fig. 2-6, the assessment degree of accuracy of first set product to TBI patient injury's orders of severity is 100%；Second
In the measurement result of set product, patients with mild quantity is reduced to 43 by 55, and moderate patient is increased to 64 by 61, weight
Degree patient is increased to 71 by 62, assesses the degree of accuracy and drops to 86.52% by 100%；The measurement result of 3rd set product
In, the patients with mild quantity in measurement result rises to 63 by 55, and moderate patient drops to 57 by 61, and severe is suffered from
Person drops to 58 by 62, assesses the degree of accuracy and drops to 88.76% by 100%；In the measurement result of 4th set product, survey
The patients with mild quantity determined in result drops to 40 by 55, and moderate patient drops to 60 by 61, and severe patient is by 62
Example rises to 78, assesses the degree of accuracy and drops to 82.02% by 100%；In the measurement result of 5th set product, measurement result
In patients with mild quantity drop to 44 by 55, moderate patient rises to 65 by 61, and severe patient is by 62 risings
To 69, assess the degree of accuracy and drop to 87.64% by 100%.
Therefore, the rabbit-anti PLG antibody of abcam companies to the sensitivity and specificity of TBI patient injury's orders of severity most
Greatly.
The antibody holding time of test example 3 tests
The anti-PLG antibody of enzyme mark is positioned in embodiment 1-3 and reference examples 1-3 kit, and kit is placed
Preserve in -20 DEG C of refrigerators, sampled behind 0,1,2,3,6,12,24 and 36 months, determine its protein content, as a result see
Table 1.
Result of the test of the different kit of table 1 to the holding time of antibody
It is that routine is carried out on the basis of kit provided by the invention by above-mentioned results showed that reference examples 1-3
Increase, delete and replace, as increased citric acid in reference examples 1, reference examples 2 replace serum with citric acid, and reference examples 3 are deleted
1640 culture medium.
Within 24 months, protein concentration does not have the protein concentration of the anti-PLG antibody of enzyme mark in embodiment 1-3 kit
Change, at 36th month, protein concentration varied slightly, and the egg of the anti-PLG antibody of the enzyme mark in reference examples 1-3 kit
White concentration even began to reduce for three month at 6th month, and during by 36th month, protein concentration is extremely low, and embodiment 2
The holding time of kit PLG antibody anti-to enzyme mark is longer than embodiment 1 and embodiment 3, it was demonstrated that kit provided by the invention
The holding time of PLG antibody anti-to enzyme mark is longer, it was demonstrated that when kit includes 3 portions of trehaloses, 25 parts of glycerine, 25 parts of blood
During cleer and peaceful 15 parts of 1640 culture medium, the holding time of PLG antibody anti-to enzyme mark is most long, and then ensures the anti-PLG antibody of enzyme mark
Specificity and sensitivity.
The dilution test of the anti-PLG antibody of the enzyme mark of test example 4
Dilute, and see in the dilution for the kit that the anti-PLG antibody of enzyme mark is positioned over into embodiment 4-6 and reference examples 4-6
The uniformity of the anti-PLG antibody of enzyme mark after diluting is examined, the results are shown in Table 2.
The anti-PLG antibody dilution test result of the enzyme mark of table 2
It is that routine is carried out on the basis of kit provided by the invention by above-mentioned results showed that reference examples 4-6
Increase, delete and replace, as increased sodium sulphate in reference examples 4, reference examples 5 replace BSA with sodium sulphate, and reference examples 6 are deleted
tween100。
From above-mentioned experimental result, the dilution PLG antibody anti-to enzyme mark in embodiment 4-6 kit carries out dilute
The uniformity after releasing is more than 90%, and the PLG antibody progress anti-to enzyme mark of the dilution in reference examples 4-6 kit is dilute
The uniformity after releasing is relatively low, and the dilution effect of the PLG antibody anti-to enzyme mark of the dilution in the kit of embodiment 5 is good
In embodiment 4 and embodiment 6, it was demonstrated that when kit includes 60 parts of PBS, tween100 and 3 of 15 parts part of BSA, to enzyme
The dilution effect for marking anti-PLG antibody is best.
Sequence table
<110>Beijing Zhen Huikang bio tech ltd
<120>A kind of anti-PLG new application and kit
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 810
<212> DNA
<213>Artificial sequence ()
<400> 1
mehkevvlll llflksgqge plddyvntqg aslfsvtkkq lgagsieeca akceedeeft 60
crafqyhske qqcvimaenr kssiiirmrd vvlfekkvyl secktgngkn yrgtmsktkn 120
gitcqkwsst sphrprfspa thpsegleen ycrnpdndpq gpwcyttdpe krydycdile 180
ceeecmhcsg enydgkiskt msglecqawd sqsphahgyi pskfpnknlk knycrnpdre 240
lrpwcfttdp nkrwelcdip rcttpppssg ptyqclkgtg enyrgnvavt vsghtcqhws 300
aqtphthnrt penfpcknld enycrnpdgk rapwchttns qvrweyckip scdsspvste 360
qlaptappel tpvvqdcyhg dgqsyrgtss ttttgkkcqs wssmtphrhq ktpenypnag 420
ltmnycrnpd adkgpwcftt dpsvrweycn lkkcsgteas vvapppvvll pdvetpseed 480
cmfgngkgyr gkrattvtgt pcqdwaaqep hrhsiftpet npraglekny crnpdgdvgg 540
pwcyttnprk lydycdvpqc aapsfdcgkp qvepkkcpgr vvggcvahph swpwqvslrt 600
rfgmhfcggt lispewvlta ahcleksprp ssykvilgah qevnlephvq eievsrlfle 660
ptrkdiallk lsspavitdk vipaclpspn yvvadrtecf itgwgetqgt fgagllkeaq 720
lpvienkvcn ryeflngrvq stelcaghla ggtdscqgds ggplvcfekd kyilqgvtsw 780
glgcarpnkp gvyvrvsrfv twiegvmrnn 810
Claims (10)
1. for detecting application of the PLG material in the product of assessment or aided assessment TBI patient injury's orders of severity is prepared.
2. material the answering in the kit of assessment or aided assessment TBI patient injury's orders of severity is prepared for detecting PLG
With.
3. the material as claimed in claim 2 for being used to detect PLG is preparing assessment or the serious journey of aided assessment TBI patient injuries
Application in the kit of degree, it is characterised in that the antibody that the material for being used to detect PLG is anti-PLG.
4. the material as claimed in claim 3 for being used to detect PLG is preparing assessment or the serious journey of aided assessment TBI patient injuries
Application in the kit of degree, it is characterised in that the antibody specific of the PLG is rabbit-anti PLG antibody.
5. the material as claimed in claim 4 for being used to detect PLG is preparing assessment or the serious journey of aided assessment TBI patient injuries
Application in the kit of degree, it is characterised in that the rabbit-anti PLG antibody is the rabbit-anti PLG antibody of abcam companies.
6. the material as claimed in claim 2 for being used to detect PLG is preparing assessment or the serious journey of aided assessment TBI patient injuries
Application in the kit of degree, it is characterised in that the sequence of the PLG is as shown in SEQ ID NO.1.
7.PLG is preparing the application in being used to assess the label of the TBI injury in traumatic condition of patient orders of severity.
A kind of 8. kit for being used for assessment or aided assessment TBI patient injury's orders of severity, it is characterised in that the kit
Including the material for detecting PLG.
9. kit as claimed in claim 8, it is characterised in that the kit also include preserve liquid, it is described preservation liquid by
Parts by weight are the trehalose of 1-5 parts, the glycerine of 15-30 parts, the serum of 20-30 parts and the 1640 culture medium of 10-20 parts composition.
10. kit as claimed in claim 8, it is characterised in that the kit also includes dilution, the dilution by
The BSA compositions of tween100 and 1-5 parts of PBS, 10-25 part of 50-70 parts.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710775306.8A CN107748263A (en) | 2017-08-31 | 2017-08-31 | The new application and kit of a kind of plasminogen |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710775306.8A CN107748263A (en) | 2017-08-31 | 2017-08-31 | The new application and kit of a kind of plasminogen |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107748263A true CN107748263A (en) | 2018-03-02 |
Family
ID=61254830
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710775306.8A Pending CN107748263A (en) | 2017-08-31 | 2017-08-31 | The new application and kit of a kind of plasminogen |
Country Status (1)
| Country | Link |
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| CN (1) | CN107748263A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112924678A (en) * | 2021-01-25 | 2021-06-08 | 四川大学华西医院 | Kit for identifying benign and malignant thyroid nodules |
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2017
- 2017-08-31 CN CN201710775306.8A patent/CN107748263A/en active Pending
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| US20090087868A1 (en) * | 2004-04-15 | 2009-04-02 | Kevin Ka-Wang Wang | Neural Proteins as Biomarkers for Nervous System Injury and Other Neural Disorders |
| US20130252834A1 (en) * | 2010-05-21 | 2013-09-26 | Universite De Geneve | Diagnostic Methods |
| CN101846682A (en) * | 2010-06-17 | 2010-09-29 | 新疆维吾尔自治区疾病预防控制中心 | Enzyme-linked immunosorbent assay sandwich kit for detecting animal plague antibody |
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| 刘红 等: "ELISA检测纤维蛋白溶酶原含量", 《临床检验杂志》 * |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112924678A (en) * | 2021-01-25 | 2021-06-08 | 四川大学华西医院 | Kit for identifying benign and malignant thyroid nodules |
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Effective date of registration: 20200131 Address after: 100032 Beijing city Xicheng District Xishiku Street No. 31 Building No. 23 hospital Applicant after: Beijing tangyihuikang Biomedical Technology Co., Ltd Address before: 100034 Beijing city Xicheng District Xishiku Street No. 31 Building No. 23 hospital 2 floor Room 202 Applicant before: BEIJING ZHEN HUIKANG BIOLOGICAL TECHNOLOGY Co.,Ltd. |
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Application publication date: 20180302 |